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17-10130 ChIPAb+™ Trimethyl-Histone H3 (Lys79) - ChIP Validated Antibody and Primer Set

This ChIPAb+ Trimethyl-Histone H3 (Lys79) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Key Applications: Western Blotting, Chromatin Immunoprecipitation (ChIP), Multiplexing Research Category: Epigenetics & Nuclear Function Research Sub-Category: Histones, Chromatin Biology
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17-10130
25 assays  25 assays per set. Recommended use: ~1 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).
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      Key Spec Table

      Species ReactivityKey Applications
      H, VrtWB, ChIP, Mplex
      Description
      Catalogue Number17-10130
      Brand Family Upstate
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+™ Trimethyl-Histone H3 (Lys79) - ChIP Validated Antibody and Primer Set
      OverviewAll ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
      The ChIPAb+ Trimethyl-Histone H3 (Lys79) set includes the Trimethyl-Histone H3 (Lys79) antibody, a negative control normal rabbit IgG, and qPCR primers which amplify a 166 bp region of human GAPDH. The Trimethyl-Histone H3 (Lys79) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Trimethyl-Histone H3 (Lys79)-associated chromatin.
      Alternate Names
      • H3K79me3
      • Histone H3 (tri methyl K79)
      • H3 histone family, member T
      • histone 3, H3
      • histone cluster 3, H3
      Background InformationHistones are highly conserved proteins that serve as the structural scaffold for the organization of nuclear DNA into chromatin. The four core histones, H2A, H2B, H3, and H4, assemble into an octamer (2 molecules of each). Subsequently, 146 base pairs of DNA are wrapped around the octamer, forming a nucleosome, the basic subunit of chromatin. Histone modifications regulate DNA transcription, repair, recombination, and replication. The most commonly studied modifications are acetylation, phosphorylation, methylation, and ubiquitination. Lysine methylation occurs in three distinct states, having either one (me1), two (me2) or three (me3) methyl groups attached to the amine group of the lysine side chain. Methylation of Lysine 79 in Histone 3 acts as a marker of inactive chromatin regions. Methylation by Dot1 methylase on lysine-79 (Lys79) of Histone H3 is criticial for transcriptional silencing, and it is thought that silencing proteins such as Sir3 function by blocking Dot1 methylation.
      References
      Product Information
      FormatAffinity Purified
      Control
      • Includes negative control normal rabbit IgG and primers specific for human GAPDH.
      PresentationAnti-Trimethyl-Histone H3 (Lys79) (rabbit polyclonal), Part No. CS204342. One vial containing 25 µg of purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4, 150 mM NaCl) with 0.05% sodium azide before the addition of 30% glycerol.
      Store at -20°C.
      Normal Rabbit IgG, Part No. PP64B. One vial containing 125 µg Rabbit IgG in 125 µL storage buffer containing 0.05% sodium azide.
      Store at -20°C.
      Control Primers, Part No. 22-004. One vial containing 75 μL of 5 μM of each primer specific for human GAPDH. Store at -20°C.
      FOR: TAC TAG CGG TTT TAC GGG CG
      REV: TCG AAC AGG AGG AGC AGA GAG CGA
      Quality LevelMQ100
      Applications
      ApplicationThis ChIPAb+ Trimethyl-Histone H3 (Lys79) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • Western Blotting
      • Chromatin Immunoprecipitation (ChIP)
      • Multiplexing
      Application NotesChromatin Immunoprecipitation:
      Representative lot data.
      Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 1 µg of normal rabbit IgG, or 1 µg of Anti-trimethyl-Histone H3 (Lys79) and the Magna ChIP™ A Kit (Cat. # 17-610).
      Successful immunoprecipitation of trimethyl-Histone H3 (Lys79) associated DNA fragments was verified by qPCR using Control Primers as a positive locus, and human β-globin as a negative locus (Figure 2). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
      Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
      Western Blot Analysis:
      Representative lot data.
      HeLa acid extract (lane 1) and recombinant Histone H3 (Cat # 14-494) (lane2) lysates were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-trimethyl Histone H3 (Lys79) (0.1 μg/mL). Proteins were visualized using a Donkey Anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system.
      Arrow indicates Histone H3 (~17 kDa) (Figure 3).
      Multiplex Immunoassay: Representative lot data.
      Specificity of interaction of anti-trimethyl histone H3 Lys79 antibody was assayed using the Luminex® multiplex immunoassay platform. Anti-trimethyl-Histone H3 (Lys79) diluted to 0.01 µg/mL concentration was incubated with a cocktail of xMAP microspheres conjugated to the following histone H3 peptides, monomethyl Lys79, unmodified Lys79, trimethyl Lys 27, trimethyl Lys 9, trimethyl Lys79 processed and detected using standard xMAP protocols (Figure 4).
      Biological Information
      ImmunogenLinear Peptide corresponding to human Histone H3 methylated at Lys79.
      EpitopeTrimethyl Lys79
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      HostRabbit
      SpecificityThis antibody recognizes Histone H3 Trimethylated on Lys79.
      Species Reactivity
      • Human
      • Vertebrates
      Species Reactivity NoteDemonstrated to react with human. Broad species reactivity is expected.
      Antibody TypePolyclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryHistones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3.
      Gene Symbol
      • HIST3H3
      • H3FT
      • H3t
      • H3T
      • H3/g
      • H3.4
      • H3/t
      Modifications
      • Methylation
      Purification MethodAffinity Purfied
      UniProt Number
      UniProt SummaryFUNCTION: Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.

      SIZE: 136 amino acids; 15508 Da

      SUBUNIT: The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA.

      SUBCELLULAR LOCATION: Nucleus.

      TISSUE SPECIFICITY: Expressed in testicular cells.

      DEVELOPMENTAL STAGE: Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.

      PTM: Acetylation is generally linked to gene activation. Acetylation on Lys-10 impairs methylation at Arg-9. Acetylation on Lys-19 and Lys-24 favors methylation at Arg-18 (By similarity). & Citrullination at Arg-9 and/or Arg-18 by PADI4 impairs methylation and represses transcription (By similarity). & Asymmetric dimethylation at Arg-18 by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 by PRMT5 is linked to gene repression (By similarity). & Methylation at Lys-5, Lys-37 and Lys-80 are linked to gene activation. Methylation at Lys-5 facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 and Lys-28 are linked to gene repression. Methylation at Lys-10 is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 and acetylation of H3 and H4. Methylation at Lys-5 and Lys-80 require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 and Lys-28 are enriched in inactive X chromosome chromatin (By similarity). & Phosphorylated at Thr-4 by GSG2/haspin during prophase and dephosphorylated during anaphase. At centromeres, specifically phosphorylated at Thr-12 from prophase to early anaphase. Phosphorylated at Ser-11 during the whole mitosis. Phosphorylation at Ser-11, which is linked to gene activation, prevents methylation at Lys-10 but facilitates acetylation of H3 and H4. Phosphorylated at Ser-29 by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation (By similarity). & Phosphorylation at 'Ser-11' is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at 'Ser-11' is important during interphase because it enables the transcription of genes following external stimulation, like stress or growth factors. Phosphorylation at 'Ser-11' is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylation at 'Ser-11' by AURKB/Aurora-B mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. & Ubiquitinated (By similarity).

      SIMILARITY: Belongs to the histone H3 family.
      Molecular Weight~ 17 kDa
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceChromatin Immunoprecipitation:
      Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 1 µg of normal rabbit IgG, or 1 µg of Anti-trimethyl-Histone H3 (Lys79) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of trimethyl-Histone H3 (Lys79) associated DNA fragments was verified by qPCR using Control Primers, (Figure 1).
      Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsStable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
      Packaging Information
      Material Size25 assays
      Material Package25 assays per set. Recommended use: ~1 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      17-10130 04053252376559

      Documentation

      ChIPAb+™ Trimethyl-Histone H3 (Lys79) - ChIP Validated Antibody and Primer Set SDS

      Title

      Safety Data Sheet (SDS) 

      ChIPAb+™ Trimethyl-Histone H3 (Lys79) - ChIP Validated Antibody and Primer Set Certificates of Analysis

      TitleLot Number
      ChIPAb+ Trimethyl-Histone H3 (Lys79) - 2051418 2051418
      ChIPAb+ Trimethyl-Histone H3 (Lys79) - NG1937491 NG1937491
      ChIPAb+ Trimethyl-Histone H3 (Lys79) - NRG1804156 NRG1804156
      ChIPAb+™ Trimethyl-Histone H3 (Lys79) - 3246318 3246318
      ChIPAb+™ Trimethyl-Histone H3 (Lys79) -2585844 2585844
      ChIPAb+™ Trimethyl-Histone H3 (Lys79) Polyclonal Antibody/Primer Set 3006559
      ChIPAb+™ Trimethyl-Histone H3 (Lys79) Polyclonal Antibody/Primer Set 2930272
      ChIPAb+™ Trimethyl-Histone H3 (Lys79) Polyclonal Antibody/Primer Set 3121623

      Brochure

      Title
      Advance your Epigenetics Research
      Product Selection Guide - Antibodies, small molecule inhibitors, kits, assays and proteins for signaling research.

      Technical Info

      Title
      Guide to Chromatin Immunoprecipitation: Critical Factors for Success

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      Categories

      Life Science Research > Antibodies and Assays > Immunoassays > Immunoprecipitation (IP) > Chromatin Immunoprecipitation (ChIP) > ChIP Validated Antibodies
      Life Science Research > Antibodies and Assays > Primary Antibodies
      Life Science Research > Protein Detection and Quantification > Immunoassays > Immunoprecipitation (IP) > Chromatin Immunoprecipitation (ChIP) > ChIP Validated Antibodies